A steroid alkaloid derivative 02F04 upregulates thymic stromal lymphopoietin expression slowly and continuously through a novel Gq/11-ROCK-ERK1/2 signaling pathway in mouse keratinocytes.
Thymic stromal lymphopoietin (TSLP), a master switch of allergic inflammation, plays an important role in the pathogenesis of allergic diseases. Although many compounds upregulate TSLP expression in vivo or in vitro, most of them are pollutants or toxicants. In the previous study, for the first time, we found that a steroid alkaloid derivative 02F04, which has a unique skeletal structure compared with other TSLP-inducing chemicals, significantly induced TSLP production in mouse keratinocytes. However, it is not investigated thoroughly that how 02F04 produces TSLP and why. In this study, we did a detailed investigation on the inducible effect and underlying molecular mechanism of 02F04 on TSLP production. We found that the peak time of TSLP mRNA level induced by 02F04 at 48 h led to a slow and continuous TSLP production in PAM212 cells. Besides, 02F04-induced TSLP production was significantly suppressed by inhibitors of Rho-associated protein kinase (ROCK), guanine nucleotide-binding protein subunit alpha q/11 (Gq/11) and extracellular signal-regulated kinase 1/2 (ERK1/2) at not only protein but also mRNA levels, and by siRNA-mediated knockdown of Gq or G11. This suggested that ROCK, Gq/11 and ERK1/2 signaling pathways were involved in 02F04-induced TSLP production. Increase in the level of p-ERK1/2 induced by 02F04 was suppressed by both inhibitors of ROCK and Gq/11, indicating that ROCK and Gq/11 molecules were located at the upstream of ERK1/2 to regulate 02F04-induced TSLP production. Gq/11 was located at the upstream of ROCK because the specific Gq/11 inhibitor of YM-254890 significantly reduced 02F04-induced actin stress fiber formation. Taken together, 02F04 upregulates a slow and continuous TSLP production through a novel Gq/11-ROCK-ERK1/2 signaling pathway. The thorough understanding the effect and mechanism of 02F04 on TSLP production is expected to supply it as a novel TSLP-regulating compound and a potential new tool for investigating the role of TSLP in allergic disorders.


Multifractal analysis of diurnal temperature range over Southern Spain using validated datasets.
Multifractal properties of diurnal temperature range (DTR) have been analyzed in this work, using validated data of maximum and minimum temperature from 197 weather stations in Southern Spain (Andalusia region). DTR is a crucial factor to characterize the regional climate, providing more information than the average daily temperature. Apart from climate change studies, one of the most important applications of DTR in Agrometeorology is as an input variable in the solar radiation or reference evapotranspiration estimation models based on the temperature. With the aim of obtaining a detailed information for different time scales, different multifractal approaches have been applied. Different quality control methods such as range/limits or persistence tests were previously applied in order to detect incorrect and anomalous values, being discarded in the subsequent analysis. The DTR scaling of moments has been analyzed and the moment scaling exponent function K(q) has been obtained, finding some differences between weather stations. In addition, multifractal dimension (D1) and multifractal degree (MD) were also estimated, revealing differences at coastal and inland locations that show heterogeneity across the region, including its multifractal nature and its invariance for a range of scales. The nonlinear characterization carried out in this work improves the understanding of DTR as an indicator of climate changes, and it can have a very positive impact on the calibration of regional models for estimating solar radiation or reference evapotranspiration based on the temperature. This multifractal characterization can be used to group stations with similar nonlinear dynamics, regardless of their geographical features, in such a way that more accurate coefficients than conventional ones are used.


Misophonia: A Review of the Literature and Its Implications for the Social Work Profession.
Misophonia is a chronic condition that describes aversion to specific auditory stimuli. Misophonia is characterized by physiological responsivity and negative emotional reactivity. Specific sounds, commonly referred to as "triggers," are often commonplace and sometimes repetitive. They include chewing, coughing, slurping, keyboard tapping, and pen clicking. Common emotional responses include rage, disgust, anxiety, and panic while physical responses include muscle constriction and increased heart rate. This literature review identifies research priorities, limitations, and new directions, examining the implications of misophonia for the social work profession. Misophonia is largely absent from the social work literature. However, the profession is uniquely equipped to understand, screen for, and effectively treat misophonia in direct practice or within interprofessional treatment teams. By conceptualizing misophonia as idiosyncratic and contextual, social workers would enhance the existing body of research by applying an ecological perspective which captures the interaction of individuals and environments in producing human experience. Such an approach would assist clients and clinicians in developing treatment plans that consider the roles of social and physical environments in the development and course of misophonia. A discussion of current limitations within the misophonia literature further emphasizes the need for new perspectives.


Effects of thyroxine binding on the stability, conformation, and fluorescence properties of thyroxine-binding globulin.
The effects of thyroxine (T4) on several molecular properties of human thyroxine-binding globulin (TBG) have been evaluated. Changes in the sedimentation constant and relaxation time indicate that TBG becomes more symmetric and compact when T4 is bound. This modification in structure is associated with an increase in the stability of TBG to denaturation by either acid or guanidinium chloride. T4 binding also produces changes in the emission and excitation spectra of TBG, reflecting different environments of the four tryptophanyl residues. T4 preferentially quenches residues in a less polar environment. In addition, it alters the effect of the collisional quencher, acrylamide, so as to indicate a shift in the environment of some of the exposed tryptophanyl residues.


RESPIRE 1: a phase III placebo-controlled randomised trial of ciprofloxacin dry powder for inhalation in non-cystic fibrosis bronchiectasis.
We evaluated the efficacy and safety of ciprofloxacin dry powder for inhalation (DPI) in patients with non-cystic fibrosis bronchiectasis, two or more exacerbations in the previous year and pre-defined bacteria in sputum.In this phase III, double-blind, placebo-controlled trial, patients were randomised 2:1 to twice-daily ciprofloxacin DPI 32.5 mg or placebo in two treatment regimens consisting of on/off treatment cycles of 14 or 28 days for 48 weeks. The primary end-points were time to first exacerbation and frequency of exacerbations.A total of 416 patients were randomised to the 14-day on/off regimen (ciprofloxacin DPI (n=137) and placebo (n=68)) or the 28-day on/off regimen (ciprofloxacin DPI (n=141) and placebo (n=70)). Ciprofloxacin DPI 14 days on/off significantly prolonged time to first exacerbation versus pooled placebo (median time >336 versus 186 days; hazard ratio 0.53, 97.5% CI 0.36-0.80; p=0.0005) and reduced the frequency of exacerbations compared with matching placebo by 39% (mean number of exacerbations 0.6 versus 1.0; incidence rate ratio 0.61, 97.5% CI 0.40-0.91; p=0.0061). Outcomes for ciprofloxacin DPI 28 days on/off were not statistically significantly different from placebo. The safety profile of ciprofloxacin DPI was favourable.Ciprofloxacin DPI was well tolerated and has the potential to be an effective treatment option in non-cystic fibrosis bronchiectasis.


Internal carotid arterial stenosis: CT angiography with volume rendering.
To determine the reliability of helical computed tomography (CT) with volume rendering for evaluation of internal carotid arterial stenosis. In 22 patients, 44 carotid arteries were evaluated with helical CT and selective conventional angiography. CT data were displayed on volume-rendered and maximum intensity projection (MIP) images. Stenoses were measured separately on axial, volume-rendered, and MIP images and on conventional angiograms. Each artery was then graded as having no stenosis, mild (< 30%) stenosis, moderate (30%-70%) stenosis, severe (> 70%) stenosis, near occlusion, or occlusion. One case of stenosis was not assessable at axial CT because of an inappropriate scanning plane; four cases were not assessable at MIP CT because of mural calcifications. All carotid arteries were assessable on volume-rendered images despite no depiction of the residual lumen at the site of narrowing in three cases of near occlusion. Correlations between angiography and helical CT were good. Axial, volume-rendered, and MIP images enabled correct classification of stenosis in 88%, 89%, and 90% of arteries, respectively. CT with volume rendering was slightly more sensitive for determining candidates for endarterectomy (i.e., those with > 70% stenosis and near occlusion); sensitivity was 100% and specificity, 92%. CT angiography with volume rendering enabled accurate evaluation of carotid disease, even when dense calcifications were present. However, no definite advantage over currently available techniques for CT measurement of stenosis severity was found.


Autoimmunity to GABAA-receptor-associated protein in stiff-person syndrome.
Stiff-person syndrome (SPS) is an autoimmune neurological disorder characterized by autoantibodies to glutamic acid decarboxylase (GAD), the enzyme responsible for the synthesis of inhibitory neurotransmitter GABA. To search for biomarkers that distinguish SPS from other neurological disorders (OND), we used surface enhanced laser desorption/ionization-time of flight (SELDI-TOF) mass spectrometry to obtain proteomic profile of sera from 25 GAD-positive SPS patients and 25 controls. A significant decrease was found in the level of a protein corresponding to GABA(A)-receptor-associated protein (GABARAP), which is responsible for the stability and surface expression of the GABA(A)-receptor. Up to 70% of the SPS sera examined, compared with 10% of the controls, immunoprecipitated GABARAP protein. Antibodies raised against GABARAP immunostained neuronal cell bodies as well as axonal and dendritic processes, as visualized by confocal microscopy. In vitro experiments demonstrated that the IgG from GABARAP antibody-positive patients, but not control IgG, significantly inhibited the surface expression of GABA(A)-receptor. We conclude that GABARAP is a new autoantigen in SPS. Because the patients' IgG inhibits the expression of GABA(A)-receptors, the circulating antibodies could impair GABAergic pathways and play a role in the clinical symptomatology of SPS patients.


Membrane-targeted self-assembling cyclic peptide nanotubes.
Peptide nanotubes are novel supramolecular nanobiomaterials that have a tubular structure. The stacking of cyclic components is one of the most promising strategies amongst the methods described in recent years for the preparation of nanotubes. This strategy allows precise control of the nanotube surface properties and the dimensions of the tube diameter. In addition, the incorporation of 3- aminocycloalkanecarboxylic acid residues in the nanotube-forming peptides allows control of the internal properties of the supramolecular tube. The research aimed at the application of membrane-interacting self-assembled cyclic peptide nanotubes (SCPNs) is summarized in this review. The cyclic peptides are designed to interact with phospholipid bilayers to induce nanotube formation. The properties and orientation of the nanotube can be tuned by tailoring the peptide sequence. Hydrophobic peptides form transmembrane pores with a hydrophilic orifice, the nature of which has been exploited to transport ions and small molecules efficiently. These synthetic ion channels are selective for alkali metal ions (Na(+), K(+) or Cs(+)) over divalent cations (Ca(2+)) or anions (Cl(-)). Unfortunately, selectivity was not achieved within the series of alkali metal ions, for which ion transport rates followed the diffusion rates in water. Amphipathic peptides form nanotubes that lie parallel to the membrane. Interestingly, nanotube formation takes place preferentially on the surface of bacterial membranes, thus making these materials suitable for the development of new antimicrobial agents.


Lithium-induced gray matter volume increase as a neural correlate of treatment response in bipolar disorder: a longitudinal brain imaging study.
Preclinical studies suggest that lithium may exert neurotrophic effects that counteract pathological processes in the brain of patients with bipolar disorder (BD). To describe and compare the course and magnitude of gray matter volume changes in patients with BD who are treated with lithium or valproic acid (VPA) compared to healthy comparison subjects, and to assess clinical relationships to gray matter volume changes induced by lithium in patients with BD, we conducted longitudinal brain imaging and clinical evaluations of treatment response in 22 mood-stabilizing and antipsychotic medications-naive patients with BD who were randomly assigned to either lithium or VPA treatment after baseline assessment. Fourteen healthy comparison subjects did not take any psychotropic medications during follow-up. Longitudinal data analyses of 93 serial magnetic resonance images revealed lithium-induced increases in gray matter volume, which peaked at week 10-12 and were maintained through 16 weeks of treatment. This increase was associated with positive clinical response. In contrast, VPA-treated patients with BD or healthy comparison subjects did not show gray matter volume changes over time. Results suggest that lithium induces sustained increases in cerebral gray matter volume in patients with BD and that these changes are related to the therapeutic efficacy of lithium.


Early profiles of axonal growth and astroglial response after spinal cord hemisection and implantation of Schwann cell-seeded guidance channels in adult rats.
We previously demonstrated that transplantation of Schwann cell-seeded channels promoted the regrowth of injured axons in the adult spinal cord. It is not clear, however, whether injured axons recapitulate the developmental scenarios to accomplish regeneration. In the present study, we investigated the early events associated with axonal regrowth after spinal cord hemisection at the eighth thoracic level and implantation of a Schwann cell-seeded minichannel in adult rats. Animals were sacrificed at postoperative days (PO) 2, 4, 7, and 14. Anterograde tracing with fluoro-ruby showed that regenerating axons grew into the graft prior to PO2 and reached the distal end of the channel at PO7. These axons expressed both embryonic neural cell adhesion molecule (E-NCAM) and growth associated protein-43 (GAP-43). Although the expression of E-NCAM decreased by PO7, that of GAP-43 remained high throughout the first 2 weeks after implantation. A close relation of vimentin-positive astroglia to the growing axons in the host tissue suggested a contact-mediated role of these cells in axon guidance. Aggregation of glial fibrillary acidic protein (GFAP)-positive astrocytes together with the increased expression of chondroitin sulfate proteoglycans (CSPGs) starting at PO7 appeared to inhibit axonal growth at the host-graft interface. Thus, adult regenerating axons and astroglia do express developmentally related molecules that may facilitate axonal growth into a permissive graft at the early phase of injury and regeneration. These results suggest that molecules and astroglia essential to development are both important in influencing axonal regrowth in the adult spinal cord.


Suppression of sprouting: An early function of NMDA receptors in the absence of AMPA/kainate receptor activity.
Electrophysiological studies have documented the existence of synapses showing only NMDA ionotropic glutamate receptor function that are therefore presumably "silent" at resting membrane potentials. Silent synapses are more prevalent in young than in older neurons, and NMDA receptor activity at such contacts may facilitate the appearance of functional AMPA receptors. However, it is uncertain whether such silent synapses actually have a function in young neurons independent of AMPA receptor induction. Using a newly characterized culture system for neurons from larval Xenopus tecta, we show that blocking NMDA receptors or preventing changes in intracellular free Ca2+ concentration with BAPTA AM significantly increases neurite sprouting and elongation in contacted but not in isolated neurons. Blocking AMPA/KA receptors or Na+-dependent action potentials does not mimic this effect. Moreover, in these young neurons, NMDA receptor-dependent Ca2+ responses to glutamate measured with confocal fluo-3 imaging are retained during AMPA/KA receptor blockade. The data suggest that many of the young contacts in these cultures are active even though they use only NMDA ionotropic glutamate receptors. Calcium influx through the NMDA receptor at these contacts seems to reduce neurite motility. This effect should lead to the accumulation of glutamatergic inputs on NMDA receptor-expressing dendrites, which could facilitate the onset of AMPA/KA receptor function and the action potential-dependent phase of synaptogenesis.


SYSTEMIC TRIPLE THERAPY IN METASTATIC HORMONE SENSITIVE PROSTATE CANCER (MHSPC).
For many years, androgen deprivation therapy (ADT) as monotherapy has been the gold standard for metastatic hormone-sensitive prostate cancer (mHSPC) treatment. Several studies have been published within the last decade demonstrating a significant survival advantage resulting from combining the treatment with standard ADT plus docetaxel or androgen receptor targeted therapy (ARTA) compared to ADT monotherapy. Recently published data of the PEACE-1 and ARASENS trials suggest that in the future, triple therapy might be a treatment option for patients with mHSPC.


Validity and reliability of the tuberculosis-related stigma scale version for Brazilian Portuguese.
Stigma associated with tuberculosis (TB) has been an object of interest in several regions of the world. The behaviour presented by patients as a result of social discrimination has contributed to delays in diagnosis and the abandonment of treatment, leading to an increase in the cases of TB and drug resistance. The identification of populations affected by stigma and its measurement can be assessed with the use of valid and reliable instruments developed or adapted to the target culture. This aim of this study was to analyse the initial psychometric properties of the Tuberculosis-Related Stigma scale in Brazil, for TB patients. The Tuberculosis-Related Stigma scale is a specific scale for measuring stigma associated with TB, originally validated in Thailand. It presents two dimensions to be assessed, namely Community perspectives toward tuberculosis and Patient perspectives toward tuberculosis. The first has 11 items regarding the behaviour of the community in relation to TB, and the second is made up of 12 items related to feelings such as fear, guilt and sorrow in coping with the disease. A pilot test was conducted with 83 TB patients, in order to obtain the initial psychometric properties of the scale in the Brazilian Portuguese version, enabling simulation of the field study. As regards its psychometric properties, the scale presented acceptable internal consistency for its dimensions, with values ≥0.70, the absence of floor and ceiling effects, which is favourable for the property of scale responsiveness, satisfactory converging validity for both dimensions, with values over 0.30 for initial studies, and diverging validity, with adjustment values different from 100%. The results found show that the Tuberculosis-Related Stigma scale can be a valid and reliable instrument for the Brazilian context.


Effects of soft contact lenses on conjunctival surface.
To examine the effects of soft contact lenses on tear breakup time (TBUT), basal Schirmer test result, and the conjunctival surface in patients wearing contact lenses. In this study, conjunctival cytologic changes, TBUT, and Schirmer test function alterations of soft contact lens wearers were evaluated by impression cytology. The study included 100 eyes of 50 soft contact lens wearers who were followed up in the Department of Ophthalmology of Dicle University and 80 eyes of 40 subjects as a control group. After TBUT and basal Schirmer test, conjunctival surface epithelial morphology was investigated using impression cytology. After the materials were appropriately stained, they were evaluated according to the Nelson grading scale. Contact lens wearers were divided into three groups according to the duration of contact lens wear. When these groups were evaluated according to the Nelson grading method, 21% of cases were grade 0; 32% were grade 1; and 28% were grade 3. There were statistically significant differences in epithelial cell morphology, goblet cell density, snakelike chromatin changes, TBUT, and basal Schirmer test result between the control and study groups. We suggest that TBUT and Schirmer test result be carefully monitored in contact lens wearers. Impression cytology may be used as a safe, simple, and noninvasive method in the diagnosis of ocular surface alternations in patients with contact lens intolerance.


Methotrexate in systemic lupus erythematosus: a systematic review of its efficacy.
The objective of this review is to evaluate the evidence for efficacy of methotrexate (MTX) in systemic lupus erythematosus (SLE). A comprehensive, computerized search was performed in MEDLINE (PubMed), EMBASE and the Cochrane Controlled Trials registry to screen for studies that examined the efficacy of MTX in adult SLE patients. The Jadad scoring system was used to assess study quality, and data were pooled using the random effects model. Of the 53 articles that were identified, 44 were excluded. Nine studies (including three randomized controlled and six observational) were eligible for inclusion. All of the included studies predominantly involved patients with arthritis or mucocutaneous features. There was significant reduction of the SLE Disease Activity Index (SLEDAI) among MTX-treated patients when compared with controls (p = 0.001, odds ratio (OR) 0.444, 95% confidence interval (CI) 0.279 to 0.707). There was also significant reduction in the average dose of corticosteroids among MTX-treated patients when compared with controls (p = 0.001, OR 0.335, 95% CI 0.202 to 0.558). The effect of MTX on laboratory and serological markers, including erythrocyte sedimentation rate, anti-dsDNA and complement levels (C3 and C4), could not be determined because of the limited numbers of controlled trials. The use of MTX is associated with significant reductions in SLEDAI and the average dose of corticosteroids in adult patients with SLE.


Interstitial cystitis pathogenesis and treatment.
Interstitial cystitis remains an idiopathic illness characterized by urinary frequency, urgency and pelvic pain with substantial morbidity in those affected. There is significant variability in the presentation, severity of symptoms and response to therapy. This review focuses on recent findings on the possible pathogenesis and potential treatments for this disease. Interstitial cystitis is manifested by sensory hypersensitivity. A small volume of urine will be associated with an exaggerated sensation of pain or pressure and urinary urgency. There is continued research regarding how this process is initiated and maintained and to what extent systemic dysfunction of the immune or autonomic nervous system may play a role. The urothelial lining has been demonstrated to be capable of secreting a large number of potential signaling molecules that may be significant factors in the disease. The pathogenesis of interstitial cystitis remains uncertain and the illness has significant diversity. Additional research is needed to establish subtypes that share common processes that can be targeted for treatment as a single effective therapy for the condition remains elusive.


Role of chemokines in promoting instability of coronary atherosclerotic plaques and the underlying molecular mechanism.
Our aim was to investigate the role of chemokines in promoting instability of coronary atherosclerotic plaques and the underlying molecular mechanism. Coronary angiography and intravascular ultrasound (IVUS) were performed in 60 stable angina pectoris (SAP) patients and 60 unstable angina pectoris (UAP) patients. The chemotactic activity of monocytes in the 2 groups of patients was examined in Transwell chambers. High-sensitivity C-reactive protein (hs-CRP), monocyte chemoattractant protein-1 (MCP-1), regulated on activation in normal T-cell expressed and secreted (RANTES), and fractalkine in serum were examined with ELISA kits, and expression of MCP-1, RANTES, and fractalkine mRNA was examined with real-time PCR. In the SAP group, 92 plaques were detected with IVUS. In the UAP group, 96 plaques were detected with IVUS. The plaques in the UAP group were mainly lipid 51.04% (49/96) and the plaques in the SAP group were mainly fibrous 52.17% (48/92). Compared with the SAP group, the plaque burden and vascular remodeling index in the UAP group were significantly greater than in the SAP group (P<0.01). Chemotactic activity and the number of mobile monocytes in the UAP group were significantly greater than in the SAP group (P<0.01). Concentrations of hs-CRP, MCP-1, RANTES, and fractalkine in the serum of the UAP group were significantly higher than in the serum of the SAP group (P<0.05 or P<0.01), and expression of MCP-1, RANTES, and fractalkine mRNA was significantly higher than in the SAP group (P<0.05). MCP-1, RANTES, and fractalkine probably promote instability of coronary atherosclerotic plaque.


Pulsar spins from an instability in the accretion shock of supernovae.
Rotation-powered radio pulsars are born with inferred initial rotation periods of order 300 ms (some as short as 20 ms) in core-collapse supernovae. In the traditional picture, this fast rotation is the result of conservation of angular momentum during the collapse of a rotating stellar core. This leads to the inevitable conclusion that pulsar spin is directly correlated with the rotation of the progenitor star. So far, however, stellar theory has not been able to explain the distribution of pulsar spins, suggesting that the birth rotation is either too slow or too fast. Here we report a robust instability of the stalled accretion shock in core-collapse supernovae that is able to generate a strong rotational flow in the vicinity of the accreting proto-neutron star. Sufficient angular momentum is deposited on the proto-neutron star to generate a final spin period consistent with observations, even beginning with spherically symmetrical initial conditions. This provides a new mechanism for the generation of neutron star spin and weakens, if not breaks, the assumed correlation between the rotational periods of supernova progenitor cores and pulsar spin.


Total pelvic exenteration with preoperative irradiation for advanced primary and recurrent rectal cancer.
To study the complication rate, local recurrence rate, and survival after total pelvic exenteration for primary advanced and recurrent rectal cancer. Prospective study. Tertiary referral university hospital, Norway. 25 patients who were operated on for primary advanced and 22 for recurrent rectal cancer since 1991; 42 men and 5 women, mean age 64 years (range 44-78). All had preoperative irradiation of 46-50 Gy. Incidence of major complications, and actuarial 5-year survival and local recurrence rate. Twenty patients had RO resection in the primary group versus seven in the recurrent group. No R2 resections were done in the primary group compared with four in the recurrent group. Half the primary cases (n = 13) had abdominoperineal resections. Hartmann's procedures were common in both groups (n = 8 in each). Postoperative mortality at 30 days was 4% (n = 2) and in-hospital 13% (n = 6). 18 patients had major complications and 12 were reoperated on. Overall 5-year actuarial survival for 43 patients without distant metastases was 28%-those with primary tumours 36%, and those with recurrent tumours 18%-similar to the figures for RO and R1 resections. Actuarial local recurrence at 5 years for primary cancers was 18% compared with 68% for recurrent cancers, again nearly identical to the figures for R0/R1 operations (p = 0.008 and p = 0.03). Some patients with advanced rectal cancer either primary or recurrent may benefit from simultaneous en-bloc cystectomy. The higher postoperative morbidity and mortality indicate the need for well-defined indications for this procedure and the necessity for thorough preoperative staging.


VIRULENCE OF MIXED-CLONE AND SINGLE-CLONE INFECTIONS OF THE RODENT MALARIA PLASMODIUM CHABAUDI.
Most evolutionary models treat virulence as an unavoidable consequence of microparasite replication and have predicted that in mixed-genotype infections, natural selection should favor higher levels of virulence than is optimal in genetically uniform infections. Increased virulence may evolve as a genetically fixed strategy, appropriate for the frequency of mixed infections in the population, or may occur as a conditional response to mixed infection, that is, a facultative strategy. Here we test whether facultative alterations in replication rates in the presence of competing genotypes occur and generate greater virulence. An important alternative, not currently incorporated in models of the evolution of virulence, is that host responses mounted against genetically diverse parasites may be more costly or less effective than those against genetically uniform parasites. If so, mixed clone infections will be more virulent for a given parasite replication rate. Two groups of mice were infected with one of two clones of Plasmodium chabaudi parasites, and three groups of mice were infected with 1:9, 5:5, or 9:1 mixtures of the same two clones. Virulence was assessed by monitoring mouse body weight and red blood cell density. Transmission stage densities were significantly higher in mixed- than in single-clone infections. Within treatment groups, transmission stage production increased with the virulence of the infection, a phenotypic correlation consistent with the genetic correlation assumed by much of the theoretical work on the evolution of virulence. Consistent with theoretical predictions of facultative alterations in virulence, we found that mice infected with both parasite clones lost more weight and had on average lower blood counts than those infected with single-clone infections. However, there was no consistent evidence of the mechanism invoked by evolutionary models that predict this effect. Replication rates and parasite densities were not always higher in ∗∗∗mixed-clone infections, and for a given replication rate or parasite density, mixed-clone infections were still more virulent. Instead, prolonged anemia and increased transmission may have occured because genetically diverse infections are less rapidly cleared by hosts. Differences in maximum weight loss occured even when there were comparable parasite densities in mixed- and single-clone infections. We suggest that mounting an immune response against more that one parasite genotype is more costly for hosts, which therefore suffer higher virulence.


Investigation of nanoyarn preparation by modified electrospinning setup.
Higher ordered structures of nanofibers, including nanofiber-based yarns and cables, have a variety of potential applications, including wearable health monitoring systems, artificial tendons, and medical sutures. In this study, twisted assemblies of polyacrylonitrile (PAN), polyvinylidene fluoride trifluoroethylene (PVDF-TrFE), and polycaprolactone (PCL) nanofibers were fabricated via a modified electrospinning setup, consisting of a rotating cone-shaped copper collector, two syringe pumps, and two high voltage power supplies. The fiber diameters and twist angles varied as a function of the rotary speed of the collector. Mechanical testing of the yarns revealed that PVDF-TrFe and PCL yarns have a higher strain-to-failure than PAN yarns, reaching 307% for PCL nanoyarns. For the first time, the porosity of nanofiber yarns was studied as a function of twist angle, showing that PAN nanoyarns are more porous than PCL yarns.


Tolerance of neural decoding errors for powered artificial legs: A pilot study.
Neural-machine interface (NMI) decoding errors challenge the clinical value of neural control of powered artificial legs, because these errors can dangerously disturb the user's walking balance, cause stumbles or falls, and thus threaten the user's confidence and safety in prosthesis use. Although extensive research efforts have been made to minimize the NMI decoding error rate, none of the current approaches can completely eliminate the errors in NMI. This study aimed at improving the robustness of prosthesis control system against neural decoding errors by introducing a fault-tolerant control (FTC) strategy. A novel reconfiguration mechanism, combined with our previously developed NMI decoding error detector, was designed and implemented into our prototypical powered knee prosthesis. The control system with FTC was preliminarily tested on two transfemoral amputees when they walked with the powered prosthesis on different walking terrains. Various NMI errors were simulated when the FTC was enabled and disabled. The preliminary testing results indicated that the FTC strategy was capable of effectively counteracting the disruptive effects of simulated decoding errors by reducing the mechanical work change around the prosthetic knee joint elicited by the NMI error. The outcomes of this study may provide a potential engineering solution to make the neural control of powered artificial legs safer to use.


Responsive eLearning exercises to enhance student interaction with metabolic pathways.
Successful learning of biochemistry requires students to engage with the material. In the past this often involved students writing out pathways by hand, and more recently directing students to online resources such as videos, songs, and animated slide presentations. However, even these latter resources do not really provide students an opportunity to engage with the material in an active fashion. As part of an online introductory metabolism course that was developed at our university, we created a series of twelve online interactive activities using Adobe Captivate 9. These activities targeted glycolysis, gluconeogenesis, the pentose phosphate pathway, glycogen metabolism, the citric acid cycle, and fatty acid oxidation. The interactive exercises consisted of two types. One involved dragging objects such as names of enzymes or allosteric modifiers to their correct drop locations such as a particular point in a metabolic pathway, a specific enzyme, and so forth. A second type involved clicking on objects, locations within a pathway, and so forth, in response to a particular question. In both types of exercises, students received feedback on their decisions in order to enhance learning. The student feedback received on these activities was very positive, and indicated that they found them to increase their confidence in the material and that they had learned the key principles of each pathway. © 2018 by The International Union of Biochemistry and Molecular Biology, 46(3):223-229, 2018.


[Structure of reciprocal connections in visual cortical areas 17 and 18 in cats].
Single cortical columns of areas 17, 18 in the cat were microiontophoretically injected with horseradish peroxidase. Spatial and laminar distributions of retrogradell labelled cells in both areas were investigated. Following injections in area 17 or in area 18 the labelled cells' region in area 17 was elongated (in a tangential plane) along the representation of visual field horizontal meridian. However the labelled cells' region in area 18 was elongated along the representation of vertical meridian. Such projection patterns appear to be common in these cortical areas throughout the central 10 degrees on various elevations (from -40 degrees to +10 degrees) of the visual field representation. Thus the spatial arrangement of intrinsic and extrinsic connections in each area coincides, at the same time in area 17 they are orthogonal to area 18. The following visual information exchange scheme may be suggested. Area 17 may supply the area 18 with more detailed information on the horizontal component of the visual image, and in the opposite direction the information on the vertical component of the same image may be supplied.


Silica sources for arsenic mitigation in rice: machine learning-based predictive modeling and risk assessment.
Arsenic (As) is a well-known human carcinogen, and the consumption of rice is the main pathway for the South Asian people. The study evaluated the impact of the amendments involving CaSiO3, SiO2 nanoparticles, silica solubilizing bacteria (SSB), and rice straw compost (RSC) on mitigation of As toxicity in rice. The translocation of As from soil to cooked rice was tracked, and the results showed that RSC and its combination with SSB were the most effective in reducing As loading in rice grain by 53.2%. To determine the risk of dietary exposure to As, the average daily intake (ADI), hazard quotient (HQ), and incremental lifetime cancer risk (ILCR) were computed. The study observed that the ADI was reduced to one-third (0.24 μg kg-1bw) under RSC+SSB treatments compared to the control. An effective prediction model was established using random forest model and described the accumulation of As by rice grains depend on bioavailable As, P, and Fe which explained 48.5, 5.07%, and 2.6% of the variation in the grain As, respectively. The model anticipates that to produce As benign rice grain, soil should have P and Fe concentration more than 30 mg kg-1 and 12 mg kg-1, respectively if soil As surpasses 2.5 mg kg-1.


Is the western scrub-jay (Aphelocoma californica) really an underdog among food-caching corvids when it comes to hippocampal volume and food caching propensity?
Food caching has been linked to better performance on spatial memory tasks and enlarged hippocampal volume in both birds and mammals. Within food-caching birds, it has also been predicted that species less reliant on stored food should have inferior spatial memory and a smaller hippocampus compared to species that depend heavily on food caches. Several comparisons suggest that North American corvids have a significantly smaller hippocampus and overall brain volume compared to the Eurasian corvid species and that western scrub-jays (Aphelocoma californica) have a smaller hippocampus compared to the more specialized Clark's nutcracker. Here we present the largest data set of scrub-jay brains and, in contrast to previous reports, show that relative to body mass western scrub-jays have a brain size similar to the largest brain size of Eurasian corvids. The relative hippocampal volume of scrub-jays is also among the largest of all investigated corvids. These findings may not be surprising considering that scrub-jays have been reported to have remarkable cognitive capacities such as episodic-like memory and experience projection. Our data suggest that many previously made assumptions about western scrub-jays as less specialized food hoarders might be an oversimplification and that simple categorization of species into specialized and non-specialized hoarders might not provide useful insights into the evolution of memory and the hippocampus.


Dermatofibrosarcoma protuberans: a comprehensive review and update on diagnosis and management.
Dermatofibrosarcoma protuberans (DFSP) is a rare superficial tumor characterized by high rates of local recurrence and low risk of metastasis. DFSP occurs most commonly on the trunk and proximal extremities, affects all races, and often develops between the second and fifth decade of life. The tumor grows slowly, typically over years. Histologically, several variants of DFSP have been described and should be well characterized to avoid misdiagnosis with other tumors. These include pigmented (Bednar tumor), myxoid, myoid, granular cell, sclerotic, atrophic DFSP, giant cell fibroblastoma, and DFSP with fibrosarcomatous areas. Of all these variants, only the DFSP with fibrosarcomatous areas is high grade, with a higher rate of local recurrence and distant metastasis. DFSP is genetically characterized by the t(17;22)(q22;q13), resulting in the fusion of alpha chain type 1 of collagen gene and platelet-derived growth factor beta gene. This translocation is present in 90% of DFSP and represents a very useful tool in the differential diagnosis of DFSP with other tumors with similar histology. The standard treatment is wide local excision with at least a 2-cm margin. However, local recurrence after apparently adequate surgical excision is well recognized. Mohs micrographic surgery would be the treatment of choice with a better cure rate and maximal conservation of tissue. When surgery is insufficient, clinical evidence has suggested that imatinib mesylate is a safe and effective treatment in DFSP, especially in cases of local advanced or metastatic disease. This article presents an overview of the state of the art in the clinicopathological management of this disease.


Blood clots. Good when you're bleeding; bad when you're not bleeding hypercoagulability and thrombophilia--overview and case report.
A critical and recurrent situation faced by the dental clinician (DC) is that of providing care to patients who may be at risk for excessive bleeding during care or post-operatively. Bleeding disorders may be due to congenital and/or acquired conditions affecting platelets and/or the coagulation process. Less often, the DC may be providing care to a patient who has an excessive clotting disorder. This paper will provide a brief overview the pathophysiology of these disorders and treatment considerations for these patients. The focus is to provide some background information for the DC so as to be better informed if a patient does present with a particular thrombotic problem. Specific details of each disorder can vessel be assessed on an individual basis pending the diagnostic category and the patient's therapy. A clinical case report will be presented.


Acronyms Use in Abdominal Wall Reconstruction: Introduction to a New Language.
The use of acronyms in medicine is widespread, aiming to simplify and condense communication. Online communication in social media platforms seems to enhance the use of acronyms, but their efficiency in message delivery may be negated by their abundance and unfamiliarity, causing more confusion than clarity. We analyzed the use of acronyms in a closed Facebook group dedicated to abdominal wall reconstruction (AWR), as the rapid recent development of this field has resulted in many new acronyms. Our aim was to classify the different acronyms and create a public reference. The International Hernia Collaboration, a hernia-related Facebook group, now communicating more than 7500 surgeons from 99 countries, was studied, by extracting acronyms used since its inception in 2012. Acronyms were categorized and interpreted, to create a small dictionary comprised of several tables. Commonly used acronyms were identified, as well as commonly used prefixes that modify the acronyms' meaning. Tables were created, classifying acronyms by their subject: 1.Anatomy2.Diseases and clinical conditions3.Techniques and materials. The use of acronyms increased in social media-based communication. Aiming to simplify the language, the inflation of terms may have achieved the opposite, by adding a multitude of unfamiliar and confusing terms. We have created a public reference for AWR-related acronyms. Limiting the liberal creation of new acronyms is recommended, especially in a rapidly changing field as AWR.


The EMPA-REG outcome study: critical appraisal and potential clinical implications.
Diabetes health care professionals have to face a study with results of incomparable success in secondary and tertiary cardiovascular disease prevention. In the past, no studies in patients with type 2 diabetes resulted to be successful in inducing an improvement of cardiovascular prognosis, no matter whether they were focused on a target, on life-style or on pharmacological intervention. On a clinical perspective, should the diabetologist's way to think about the anti-diabetic therapy of patients on secondary cardiovascular prevention change based on the results of Empa-Reg outcome? Due to the complexity of the clinical picture of patients with type 2 diabetes, a tailored therapy based on targets, complications, co-morbidity, familial and social environment, personal and cultural features must be conceived and applied in starting pharmacological therapy; however, the question whether should we consider empagliflozin as first choice therapy in individuals with type 2 diabetes exposed to high cardiovascular risk, the Empa-Reg outcome-like patient, awaits now for an answer. Waiting for data confirming the results of the Empa-Reg outcome study, this report goes through the good reasons in support of this way of thinking, but at the same time explores the many unanswered questions raising potential concerns about this clinical choice.


Signal-to-noise ratio, contrast-to-noise ratio and their trade-offs with resolution in axial-shear strain elastography.
In axial-shear strain elastography, the local axial-shear strain resulting from the application of quasi-static axial compression to an inhomogeneous material is imaged. In this paper, we investigated the image quality of the axial-shear strain estimates in terms of the signal-to-noise ratio (SNR(asse)) and contrast-to-noise ratio (CNR(asse)) using simulations and experiments. Specifically, we investigated the influence of the system parameters (beamwidth, transducer element pitch and bandwidth), signal processing parameters (correlation window length and axial window shift) and mechanical parameters (Young's modulus contrast, applied axial strain) on the SNR(asse) and CNR(asse). The results of the study show that the CNR(asse) (SNR(asse)) is maximum for axial-shear strain values in the range of 0.005-0.03. For the inclusion/background modulus contrast range considered in this study (<10), the CNR(asse) (SNR(asse)) is maximum for applied axial compressive strain values in the range of 0.005%-0.03%. This suggests that the RF data acquired during axial elastography can be used to obtain axial-shear strain elastograms, since this range is typically used in axial elastography as well. The CNR(asse) (SNR(asse)) remains almost constant with an increase in the beamwidth while it increases as the pitch increases. As expected, the axial shift had only a weak influence on the CNR(asse) (SNR(asse)) of the axial-shear strain estimates. We observed that the differential estimates of the axial-shear strain involve a trade-off between the CNR(asse) (SNR(asse)) and the spatial resolution only with respect to pitch and not with respect to signal processing parameters. Simulation studies were performed to confirm such an observation. The results demonstrate a trade-off between CNR(asse) and the resolution with respect to pitch.


Pyrenecarboxaldehyde encapsulated porous TiO2 nanoreactors for monitoring cellular GSH levels.
Recently, ternary electrochemiluminescence (ECL) system has become a hot research topic due to its great potential for improving ECL efficiency by promoting the generation of intermediates. However, it is still a great challenge to increase the utilization rate of intermediates in a ternary ECL system. Herein, we propose a strategy to increase the utilization rate of intermediates by designing pyrenecarboxaldehyde (Pyc) encapsulated porous titania (pTiO2) nanospheres (Pyc@pTiO2) as ECL nanoreactors for an integrated ternary (luminophore/coreactant/co-reaction accelerator, Pyc/S2O82-/TiO2) ECL system construction. Specifically, pTiO2 acted as an ECL co-reaction accelerator, in which Pyc could obtain electrons from the conduction band of TiO2 to produce more SO4˙-, increasing its emissions. Simultaneously, pTiO2 could provide confined reaction spaces to effectively shorten the diffusion distance, extend the lifetime of free radicals, increase the utilization rate of intermediates and improve the efficiency of the ternary ECL system. As a proof of concept, the Pyc@pTiO2 nanoreactors-based sensing platform was successfully constructed to sensitively monitor cellular GSH levels. Overall, this work for the first time proposed an avenue to increase the utilization rate of intermediates in a ternary ECL system, which opened a new route for ECL biosensing in cell analysis applications.


Comparison of Serum PCSK9 Levels in Subjects with Normoglycemia, Impaired Fasting Glucose, and Impaired Glucose Tolerance.
We investigated proprotein convertase subtilisin/kexin type 9 (PCSK9) concentrations in individuals with normoglycemia, impaired fasting glucose (IFG), and impaired glucose tolerance (IGT). This was a pilot, cross-sectional study including 92 individuals who had not been diagnosed with or treated for diabetes. We measured PCSK9 levels in three groups of subjects; namely, normoglycemia (n=57), IFG (n=21), and IGT (n=14). Individuals with IFG and IGT showed higher PCSK9 concentrations than those in the normoglycemic group, with the highest serum PCSK9 concentrations found in individuals with IGT (55.25±15.29 ng/mL for normoglycemia, 63.47±17.78 ng/mL for IFG, 72.22±15.46 ng/mL for IGT, analysis of variance P=0.001). There were no significant differences in high- or low-density lipoprotein cholesterol among groups. Serum PCSK9 levels are increased in patients with prediabetes compared to subjects with normoglycemia.


Agar films containing silver nanoparticles as new supports for electromembrane extraction.
A new support containing silver nanoparticles to assist electromembrane extraction (EME) procedures is proposed. For the first time, synthesized agar films containing silver nanoparticles (AgNPs) have been used as a support for liquid membranes in EME. Agarose films of 20 μm thickness containing 107.9 mg Ag/g agar were synthesized and characterized by transmission electron microscopy (TEM) and atomic force microscopy (AFM), showing isolated spherical silver nanoparticles of 20-30 nm diameter with homogeneous distribution. Nanometallic films were cut and glued to narrow bore glass tubes and used as supports for a dihexyl ether liquid membrane for use in an EME procedure. EME conditions were optimized and applied to the extraction of selected non-steroidal anti-inflammatory drugs (NSAIDs). The results were compared to those using polypropylene membranes (450 μm and 100 μm thickness), achieving 10- to 70-fold higher extraction efficiency. This article opens a new line of research into electrically assisted microextraction systems by combining other possible kinds of nanometallic films, including different metals, film functionalization through metallic NPs, and the use of low polarity solvents. Also, very low currents are obtained during the extraction process, which lead to high electromigration of the analytes.


Spatial Vertical Directionality and Correlation of Low-Frequency Ambient Noise in Deep Ocean Direct-Arrival Zones.
Wind-driven and distant shipping noise sources contribute to the total noise field in the deep ocean direct-arrival zones. Wind-driven and distant shipping noise sources may significantly and simultaneously affect the spatial characteristics of the total noise field to some extent. In this work, a ray approach and parabolic equation solution method were jointly utilized to model the low-frequency ambient noise field in a range-dependent deep ocean environment by considering their calculation accuracy and efficiency in near-field wind-driven and far-field distant shipping noise fields. The reanalysis databases of National Center of Environment Prediction (NCEP) and Volunteer Observation System (VOS) were used to model the ambient noise source intensity and distribution. Spatial vertical directionality and correlation were analyzed in three scenarios that correspond to three wind speed conditions. The noise field was dominated by distant shipping noise sources when the wind speed was less than 3 m/s, and then the spatial vertical directionality and vertical correlation of the total noise field were nearly consistent with those of distant shipping noise field. The total noise field was completely dominated by near field wind generated noise sources when the wind speed was greater than 12 m/s at 150 Hz, and then the spatial vertical correlation coefficient and directionality pattern of the total noise field was approximately consistent with that of the wind-driven noise field. The spatial characteristics of the total noise field for wind speeds between 3 m/s and 12 m/s were the weighted results of wind-driven and distant shipping noise fields. Furthermore, the spatial characteristics of low-frequency ambient noise field were compared with the classical Cron/Sherman deep water noise field coherence function. Simulation results with the described modeling method showed good agreement with the experimental measurement results based on the vertical line array deployed near the bottom in deep ocean direct-arrival zones.


Enhancement of the dimethylnitrosamine acute effects in rat liver by prior treatment with triton WR-1339.
Prior administration of Triton WR-1339 (tyloxapol, an anionic surfactant) to noninbred Sprague-Dawley male rats significantly enhanced the intensity of the necrogenic effect of dimethylnitrosamine (DMN) on the liver. This phenomenon was established by determination of NADP+-linked isocitrate dehydrogenase activity in the plasma and by histologic procedures. This enhancing effect was not due to an increase in the levels of DMN that reached the liver, because the content of DMN in the livers of Triton WR-1339-treated or untreated animals at 1 or 3 hours was not significantly different. Triton WR-1339 administration had no effect on DMN liver metabolism to formaldehyde or CO2; in addition, the covalent binding of DMN metabolites to nucleic acids or proteins was not modified by pretreatment with Triton WR-1339. However, in vitro, high concentrations (1 mg/ml) of Triton WR-1339 decreased the intensity of these parameters. This effect disappeared when the concentration was lowered to 0.4 mg/ml. Results are compatible with the hypothesis that the potentiating effects of Triton WR-1339 on liver damage caused by DMN and other hepatotoxins were due to a modification of the response of liver cells to injury.


Psychosocial factors, female fertility and pregnancy: a prospective study--Part I: Fertility.
The aim of the study was to determine prospectively whether psychosocial, personality-related or stress factors have influence on fertility. Initially 191 healthy nulliparas without a history of infertility who were planning to have children were studied gynecologically and by psychiatric examination. Life changes and changes in psychosocial stress were monitored throughout the study period of 6 months at regular intervals. The following factors turned out to be associated with higher than average fertility in the final follow-up sample of 180 women: looking younger than one's actual age, no fluctuation in body weight before pregnancy, low consumption of coffee, low score of psychosomatic symptoms, being the youngest sibling, low number of negative life changes, younger than spouse, having phobic traits, and customarily religious. There was no clearcut association between low fertility and deviations in personality factors.


Deciphering regulatory variation of THI genes in alcoholic fermentation indicate an impact of Thi3p on PDC1 expression.
Thiamine availability is involved in glycolytic flux and fermentation efficiency. A deficiency of this vitamin may be responsible for sluggish fermentations in wine making. Therefore, both thiamine uptake and de novo synthesis could have key roles in fermentation processes. Thiamine biosynthesis is regulated in response to thiamine availability and is coordinated by the thiamine sensor Thi3p, which activates Pdc2p and Thi2p. We used a genetic approach to identify quantitative trait loci (QTLs) in wine yeast and we discovered that a set of thiamine genes displayed expression-QTL on a common locus, which contains the thiamine regulator THI3. We deciphered here the source of these regulatory variations of the THI and PDC genes. We showed that alteration of THI3 results in reduced expression of the genes involved in thiamine biosynthesis (THI11/12/13 and THI74) and increased expression of the pyruvate decarboxylase gene PDC1. Functional analysis of the allelic effect of THI3 confirmed the control of the THI and PDC1 genes. We observed, however, only a small effect of the THI3 on fermentation kinetics. We demonstrated that the expression levels of several THI genes are correlated with fermentation rate, suggesting that decarboxylation activity could drive gene expression through a modulation of thiamine content. Our data also reveals a new role of Thi3p in the regulation of the main pyruvate decarboxylase gene, PDC1. This highlights a switch from PDC1 to PDC5 gene expression during thiamine deficiency, which may improve the thiamine affinity or conservation during the enzymatic reaction. In addition, we observed that the lab allele of THI3 and of the thiamin transporter THI7 have diverged from the original alleles, consistent with an adaptation of lab strains to rich media containing an excess of thiamine.


Positive effect of strontium-oxide layer on the osseointegration of moderately rough titanium surface in non-osteoporotic rabbits.
To evaluate the effect of strontium-oxide layer on new bone formation and osseointegration of sandblasted large-grit double-acid-etched (SLA) implant. Strontium-oxide layer on the SLA surface was produced by hydrothermal treatment using a Sr-containing solution. The surface topographies, roughness, hardness values, chemical elements and ionic release of SLA and the strontium-containing SLA (Sr-SLA) surface were measured by special instruments separately. Sixty-four SLA and Sr-SLA implants were inserted into the proximal tibiae and femoral condyles of sixteen non-osteoporotic New Zealand white rabbits. The biological effects were evaluated by removal torque (RTQ) testing and histomorphometric analysis after 3 and 6 weeks of implantation. The surface characteristics showed Sr-SLA surfaces with dotted nanostructures can release appropriate amount of strontium ions into surrounding tissue till 14 days. In vivo, the Sr-SLA implants presented significantly higher RTQ than SLA implants at 3 and 6 weeks (P < 0.05). The Sr-SLA implants presented higher bone-to-implant contact (BIC) than SLA implants in cortical bone at 3 and 6 weeks (P < 0.05). The bone area was slightly higher for the Sr-SLA implants at 3 and 6 weeks (P > 0.05). The strontium-oxide layer on the SLA surface has the potential to improve implant osseointegration in non-osteoporotic rabbits.


Ultrahigh sensitive and selective detection of single nucleotide polymorphism using peptide nucleic acid and ribonuclease H assembled DNA amplification (PRADA).
Early diagnosis and monitoring of cancer is the best way to increase the survival rate among patients with cancer. However, the current cancer screening technology is expensive and time-consuming; hence, cancer screening remains challenging. Therefore, developing a relatively inexpensive and high-performance analytical method is necessary. Especially, mutations in KRAS can be characterized as single nucleotide polymorphism mutations. Therefore, discrimination of single nucleotide polymorphism is essential to detect cancer mutations. This study introduces a method with high sensitivity and selectivity of real-time PCR using peptide nucleic acid (PNA) and RNase H II to detect KRAS single nucleotide polymorphism. This method was developed via the fusion of the existing PNA clamping PCR technique and the RNase H-dependent PCR technique. A synergistic effect was realized by mitigating the shortcomings of each method. Our method had a detection limit of 1 aM and selectivity of 0.01%. This study demonstrated completed validation tests with DNA-spiked plasma sample analysis, cell culture, and analysis of blood samples collected from patients with cancer. Furthermore, we demonstrated the applicability of this method for breath biopsy.


Genetic variation of contact dermatitis in broilers.
This study aimed to investigate the presence of genetic variation in footpad dermatitis (FPD) and hock burns (HB) and the possibility to genetically select against these. A field trial including 10 commercial broiler lines (n = 102 to 265) was carried out at 2 Dutch farms. Footpad dermatitis and HB were subjectively scored at approximately 4, 5, and 7 wk on a scale from 0 through 5. Genetic parameters were estimated in 2 lines based on a larger data set. The overall agreement of repeated FPD and HB scores was high (0.66 to 0.86) and the scoring system was, therefore, considered reliable. Kendall's tau between left and right scores was lower than 1 (FPD: 0.73 and HB: 0.57), and both left and right FPD and HB must, therefore, be evaluated. High prevalences of FPD, but also HB, were achieved in the field trial, but lower prevalences may be sufficient for genetic evaluations and would be less detrimental to welfare. Genetic variation between and within lines was present for both FPD and HB as indicated by between-line differences and heritabilities, and selection against FPD and HB is, therefore, possible. It is important that selection is done against both FPD and HB, and such selection should not have a negative influence on the genetic improvement in BW. In contrast, continued selection for increased BW while ignoring FPD in the breeding goal is likely to lead to an increased propensity to develop FPD in broilers.


[Complexity: concept and challenges for public health interventions].
Introduction: Since 2000, the notion of "complex interventions" has been emerging in the health research field. "Complex interventions" and "complexity" are commonly used terms, but they are generally not defined. Conceptual ambiguities persist concerning the notion of complexity. The objective of this exploratory review is to characterize the notion of complexity: What is complexity? Where does this notion come from and what does it cover? What are the consequences of complexity in the health field?Methods: To clarify the concept of complexity, a narrative review was conducted in the fields of humanities and social science, managerial economics, psychology and healthcare.Results: The concept of complexity, that can be attributed to Edgar Morin, has been the subject of appropriations, adaptations, and operations in multiple areas. Complexity consists of understanding the factors influencing individual decisions. In the field of healthcare, the concept of complexity is used more pragmatically and is defined by objective characteristics of interventions (defined as complex) or their contexts for the practical purposes of evaluation.Discussion: The notions of complexity and complex interventions have implications for researchers and users of the results of research. In particular, the notion of complexity is designed to provide a better understanding of the mechanism of effectiveness of interventions, support transferability and use by actors and decision-makers.


[Acaricidal activity of "Medifox" against the human itch mites Sarcoptes scabiei].
The acaricidal activity of Medifox was evaluated in terms of the itch mites to lend support to the validity of its regimen to treat human scabies. Experiments using mainly females, eggs, and larvae withdrawn from the burrows on the skin of patients with scabies were performed. Permethrin contained in 0.4% aqueous Medifox emulsion was found to affect an itch mite by contact and intestinal routes, without a marked knock-down effect; however, the signs of intoxication are irreversible and lead to the death of Medifox-treated mites. In an in vitro experiment, 100% death of specimens occurred 24-48 hours after immersion of the active stages in the aqueous emulsion for 1 min, depending on the stage of their development. Visual inspection of the course of embryogenesis and the hatch of larvae from eggs under a light microscope established that 20-40-min exposure to Medifox emulsion did not prevent the final formation of a larva and its hatch if the egg was at the late stages of embryogenesis. Long-term (180-min) exposure yielded a 100% ovicidal effect. An in vivo experiment indicated that there were no viable mites in the scabies burrows after single-to-double treatment of patients. Repeated applications are required due to their incomplete ovicidal effect. As a matter of fact, the third treatment is additional to create the "safety factor" of therapy. It is concluded that there is an agreement of the results of in vitro and in vivo experiments, by studying Medifox as an example. The findings suggest that the chosen Medifox treatment regimen including three consecutive treatments results in complete eradication of all the stages of the causative agent on the patient.


Aggressive pharmacologic and surgical treatment of spinal cord injuries in dogs and cats.
Aggressive pharmacologic and surgical treatment of spinal cord injuries in 22 dogs and 3 cats resulted in a recovery rate of 44% of all 25 animals. Excluding 5 animals that were euthanatized at surgery because of visualized cord transection and 5 that died during surgery because of secondary lesions, the recovery rate was 73%. This is compared with a recovery rate of 39% (7/18) for animals treated with more conservative methods at the same institution during an earlier period. Of the 7 animals without spinal cord transection and treated within 4 hours of injury, all recovered, pointing to the merits of early surgical intervention.


Effect of niuche-shen-qi-wan on painful muscle cramps in patients with liver cirrhosis: a preliminary report.
Twelve patients with liver cirrhosis complaining of painful muscle cramps were treated with Niuche-Shen-Qi-Wan (TJ-107). Three patients were at the decompensated state. Muscle cramps disappeared in 4 weeks on the average after oral administration of TJ-107 in all 12 patients. During the period of TJ-107 administration, there was no significant improvement of hepatic function. One patient complained of mild epigastric discomfort after taking TJ-107, but there were no other adverse effects. Our results indicate that TJ-107 is useful for treatment of painful muscle cramps in cirrhotic patients.


Relative roles of theca and granulosa cells in ovarian follicular steroidogenesis in the amphibian, Rana nigromaculata.
Using frog ovarian follicles in vitro we assessed the role of the theca/epithelium (THEP) layer and granulosa cells in the production of steroids. The THEP layer and granulosa cell-enclosed oocytes (GCEOs) were separated from ovarian follicles of the frog, Rana nigromaculata, by microdissection. Intact follicles (IFs), THEP layer, or GCEOs were cultured for 6 hr in amphibian Ringer's in the presence or absence of various steroid precursors (100 ng/ml) or of frog pituitary homogenate (FPH, 0.05 gland/ml). The amounts of progesterone (P4), 17 alpha-hydroxyprogesterone (17 alpha-OHP4), androstenedione (AD), testosterone (T), and estradiol (E2) secreted into the medium by the different types of follicular components during culture were measured by radioimmunoassay. Addition of pregnenolone resulted in a marked increase in P4 concentrations produced by GCEOs (1858 pg/follicle) and IFs (1549 pg/follicle), but a much smaller increase in P4 occurred in the THEP layer (427 pg/follicle). Likewise, addition of P4 stimulated greater production of 17 alpha-OHP4 by GCEOs (659 pg/follicle) and by IFs (877 pg/follicle) than by the THEP layer (330 pg/follicle). Exogenous 17 alpha-OHP4 markedly increased AD levels secreted by GCEOs (1139 pg/follicle) and by IFs (841 pg/follicle), but not by the THEP layer (< 15 pg/follicle). In contrast, AD addition resulted in the secretion of markedly elevated levels of T by the THEP layer (2326 pg/follicle) and IFs (3652 pg/follicle) as compared to the GCEOs (408 pg/follicle). Addition of T markedly increased E2 concentrations produced by GCEOs (378 pg/follicle) and by IFs (673 pg/follicle), but not by the THEP layer (< 12 pg/follicle).(ABSTRACT TRUNCATED AT 250 WORDS)


Identification of functionally important regions of the muscular chloride channel CIC-1 by analysis of recessive and dominant myotonic mutations.
Mutations in the muscular voltage-dependent Cl-channel, CIC-1, lead to recessive and dominant myotonia. Here we analyse the effects of one dominant (G200R) and three recessive (Y150C, Y261C, and M485V) mutations after functional expression in Xenopus oocytes. Glycine 200 is a highly conserved amino acid located in a conserved stretch in the putatively cytoplasmic loop between domains D2 and D3. Similar to several other dominant mutations the amino acid exchange G200R leads to a drastic shift by approximately 65 mV of the open probability curve to more positive voltages. As explored by co-expression studies, the shift is intermediate in heteromeric mutant/WT channels. Open channel properties such as single channel conductance, rectification or ion selectivity are not changed. Thus we identified a new region of the CIC-1 protein in which mutations can lead to drastic shifts of the voltage dependence. The recessive mutation M485V, which is located in a conserved region at the beginning of domain D10, leads to a drastic reduction of the single channel conductance from 1.5 pS for WT to approximately 0.3 pS. In addition, the mutant is strongly inwardly rectifying and deactivates incompletely at negative voltages. Ion-selectivity, however, is unchanged. These electrophysiological properties fully explain the recessive phenotype of the mutation and identify a new region of the protein that is involved in ion permeation and gating of the CIC-1 channel. The other two recessive mutations (Y150C and Y261C) had been found in a compound heterozygous patient. Surprisingly, expression of these mutants in oocytes yielded currents indistinguishable from WT CIC-1 when explored by two-electrode voltage clamp recording and patch clamping (either singly or both mutations co-expressed). Other mechanisms that are not faithfully represented by the Xenopus expression system must therefore be responsible for the myotonic symptoms associated with these mutations.


Functional hyperspectral imaging captures subtle details of cell metabolism in olfactory neurosphere cells, disease-specific models of neurodegenerative disorders.
Hyperspectral imaging uses spectral and spatial image information for target detection and classification. In this work hyperspectral autofluorescence imaging was applied to patient olfactory neurosphere-derived cells, a cell model of a human metabolic disease MELAS (mitochondrial myopathy, encephalomyopathy, lactic acidosis, stroke-like syndrome). By using an endogenous source of contrast subtle metabolic variations have been detected between living cells in their full morphological context which made it possible to distinguish healthy from diseased cells before and after therapy. Cellular maps of native fluorophores, flavins, bound and free NADH and retinoids unveiled subtle metabolic signatures and helped uncover significant cell subpopulations, in particular a subpopulation with compromised mitochondrial function. Taken together, our results demonstrate that multispectral spectral imaging provides a new non-invasive method to investigate neurodegenerative and other disease models, and it paves the way for novel cellular characterisation in health, disease and during treatment, with proper account of intrinsic cellular heterogeneity.


Estrogen exposure affects the post-ovariectomy increases in both LH and FSH release in female rats.
In female rats ovariectomy (OVX) on the morning of diestrus day 2 resulted in a prompt (within 8 h) and gradual increase in LH release, whereas a similar operation on estrus did not raise serum LH until 24 h later. In proestrous females, OVX in the morning neither prevented the anticipated LH surge on that evening, nor increased LH release on the next morning. Since circulating concentrations of estradiol (E2) are progressively increased during diestrus day 2 and proestrus, this pattern of E2 secretion may affect the acute increase in LH secretion following OVX. To test such a hypothesis, we examined the effects of large, sustained or large, transient increases in circulating estrogen on the subsequent increase in gonadotropin secretion after OVX. On the morning of diestrus day 1 a subcutaneous injection of 20 micrograms estradiol benzoate (EB) produced prompt and large increases in both serum E2 and estrone (E1) for about 2 days, despite OVX of females at 24 h after the EB injection. An injection of 20 micrograms 17 beta-estradiol (E2) also promptly increased both serum E2 and E1. micrograms E2 injection were twice that observed after 20 micrograms EB, and both E2 and E1 returned to low baseline values within 24 h after the E2 injection. In contrast, administration of 1 microgram EB only produced small and transient rises in serum E2 and E1.(ABSTRACT TRUNCATED AT 250 WORDS)


MiR-646 suppresses proliferation and metastasis of non-small cell lung cancer by repressing FGF2 and CCND2.
MicroRNA-646 (miR-646) has been implicated in several other cancers; however, its functional mechanism in non-small cell lung cancer (NSCLC) remains unclear. In this study, we observed the downregulation of miR-646 expression in NSCLC tissues and cell lines. Low-level expression of miR-646 was associated with metastasis and stage of NSCLCs. Functional assays showed that overexpression of miR-646 could suppress NSCLC cell proliferation, clonogenicity, invasion, and inhibit epithelial-mesenchymal transition (EMT), whereas decreased miR-646 expression showed the opposite effects. Importantly, miR-646 overexpression attenuated in vivo tumor growth and metastasis in nude mice models. Mechanically, miR-646 directly targeted and suppressed fibroblast growth factor 2 (FGF2) and cyclin D2 (CCND2) expression. Reintroduction of FGF2 and CCND2 attenuated miR-646-mediated suppression of proliferation and invasion in NSCLC. Collectively, these results demonstrate that miR-646 acts as a tumor suppressor in NSCLC by targeting FGF2 and CCND2, and may serve as a therapeutic target for patients with NSCLC.


Genome-specific repetitive DNA and RAPD markers for genome identification in Elymus and Hordelymus.
We have developed RFLP and RAPD markers specific for the genomes involved in the evolution of Elymus species, i.e., the St, Y, H, P, and W genomes. Two P genome specific repetitive DNA sequences, pAgc1 (350 bp) and pAgc30 (458 bp), and three W genome specific sequences, pAuv3 (221 bp), pAuv7 (200 bp), and pAuv13 (207 bp), were isolated from the genomes of Agropyron cristatum and Australopyrum velutinum, respectively. Attempts to find Y genome specific sequences were not successful. Primary-structure analysis demonstrated that pAgc1 (P genome) and pAgc30 (P genome) share 81% similarity over a 227-bp stretch. The three W genome specific sequences were also highly homologous. Sequence comparison analysis revealed no homology to sequences in the EMBL-GenBank databases. Three to four genome-specific RAPD markers were found for each of the five genomes. Genome-specific bands were cloned and demonstrated to be mainly low-copy sequences present in various Triticeae species. The RFLP and RAPD markers obtained, together with the previously described H and St genome specific clones pHch2 and pP1Taq2.5 and the Ns genome specific RAPD markers were used to investigate the genomic composition of a few Elymus species and Hordelymus europaeus, whose genome formulas were unknown. Our results demonstrate that only three of eight Elymus species examined (the tetraploid species Elymus grandis and the hexaploid species Elymus caesifolius and Elymus borianus) really belong to Elymus.


Metabolomic changes during cellular transformation monitored by metabolite-metabolite correlation analysis and correlated with gene expression.
To investigate metabolic changes during cellular transformation, we used a 1H NMR based metabolite-metabolite correlation analysis (MMCA) method, which permits analysis of homeostatic mechanisms in cells at the steady state, in an inducible cell transformation model. Transcriptomic data were used to further explain the results. Transformed cells showed many more metabolite-metabolite correlations than control cells. Some had intuitively plausible explanations: a shift from glycolysis to amino acid oxidation after transformation was accompanied by a strongly positive correlation between glucose and glutamine and a strongly negative one between lactate and glutamate; there were also many correlations between the branched chain amino acids and the aromatic amino acids. Others remain puzzling: after transformation strong positive correlations developed between choline and a group of five amino acids, whereas the same amino acids showed negative correlations with phosphocholine, a membrane phospholipid precursor. MMCA in conjunction with transcriptome analysis has opened a new window into the metabolome.


Macrophage-derived IL-18 targeting for the treatment of Crohn's disease.
Crohn's disease is an inflammatory bowel disease associated with several changes in the immune system, including an increased number of infiltrating macrophages. These macrophages release a variety of proinflammatory cytokines, such as tumor necrosis factor-alpha (TNF-alpha, macrophage infiltrating factor (MIF), interleukin (IL)-6, IL-12 and IL-18, which are critically involved in the onset and the development of Crohn's disease. We here focus on the role of macrophages, especially macrophage-derived IL-18 in both patients with Crohn's disease and a murine model of Crohn's disease.


MDMA and serotonin: based on two cases.
3,4-methylenedioxyamphetamine (MDMA) or "ecstasy" damages serotonin neurons in all animal species and there is growing evidence that this finding also applies to humans. This fact, together with the increasing extended use in the young population, has important repercussions in the appearance of specific psychopathologic and cognitive disturbances associated to its use. The authors present two clinical cases, in which psychopathological and cognitive symptoms are detected in MDMA users that support this hypothesis. Problems in the diagnosis of psychiatric disorders associated to MDMA and its clinical and therapeutic implications are discussed.


Participation of NO signaling in formation of long-term memory in salivary conditioning of the cockroach.
The molecular and neural basis of protein synthesis-dependent long-term memory (LTM) has been the subject of extensive studies in vertebrates and invertebrates. In crickets and honey bees, it has been demonstrated that nitric oxide (NO) signaling plays critical roles in LTM formation, but no experimental system appropriate for electrophysiological study of neural mechanisms by which production of NO leads to LTM formation has been established in insects. We have reported that cockroaches, as do dogs and humans, exhibit conditioning of salivation, i.e., they exhibit an increased level of salivation in response to an odor paired with sucrose reward. Salivary conditioning can be monitored by activity changes of salivary neurons in rigidly immobilized animals and thus is useful for the study of brain mechanisms of learning and memory. In the present study, we found that injection of cycloheximide, a protein synthesis inhibitor, into the hemolymph before multiple conditioning trials impairs formation of 1-day memory, but not that of 30-min memory. This indicates that formation of 1-day memory requires protein synthesis but that of earlier memory does not. We also found that injection of l-NAME, an inhibitor of NO synthase, before multiple conditioning impairs formation of 1-day memory but not that of 30-min memory. We thus conclude that NO signaling participates in the formation of protein synthesis-dependent LTM but not that of earlier memory in salivary conditioning. Salivary conditioning in cockroaches should become a pertinent system for the study of neural mechanisms by which activation of NO synthase leads to LTM formation.


Generation of the pitch moment during the controlled flight after takeoff of fruitflies.
In the present paper, the controlled flight of fruitflies after voluntary takeoff is studied. Wing and body kinematics of the insects after takeoff are measured using high-speed video techniques, and the aerodynamic force and moment are calculated by the computational fluid dynamics method based on the measured data. How the control moments are generated is analyzed by correlating the computed moments with the wing kinematics. A fruit-fly has a large pitch-up angular velocity owing to the takeoff jump and the fly controls its body attitude by producing pitching moments. It is found that the pitching moment is produced by changes in both the aerodynamic force and the moment arm. The change in the aerodynamic force is mainly due to the change in angle of attack. The change in the moment arm is mainly due to the change in the mean stroke angle and deviation angle, and the deviation angle plays a more important role than the mean stroke angle in changing the moment arm (note that change in deviation angle implies variation in the position of the aerodynamic stroke plane with respect to the anatomical stroke plane). This is unlike the case of fruitflies correcting pitch perturbations in steady free flight, where they produce pitching moment mainly by changes in mean stroke angle.


Draft Genome Sequence of Erysipelothrix rhusiopathiae, Isolated from a Canine Case of Diskospondylitis.
This is the draft genome of an Erysipelothrix rhusiopathiae strain isolated from the blood of a canine. Initial 16S ribosomal DNA amplification identified the isolate as belonging to the Erysipelothrix genus but could not elucidate the species due to previous misidentification of E. rhusiopathiae and E. tonsillarum The species identification was confirmed by whole-genome sequencing.


Inverse ratio ventilation (I/E = 2/1) in acute respiratory distress syndrome: a six-hour controlled study.
To assess the cardiorespiratory effects of a prolonged application of inverse ratio ventilation (IRV), we compared IRV (I/E = 2) with conventional ventilation (CV) (I/E = 0.5), applied for 6 h each in a randomized order, with constant tidal volume (VT) and total positive end-expiratory pressure (PEEP(tot)) in eight patients with acute respiratory distress syndrome (ARDS). After 1 h, IRV resulted in a lower peak inspiratory pressure (PIP) (28.2 +/- 1.5 versus 35.6 +/- 1.7 cm H2O, p < 0.05), an unchanged plateau pressure, and a higher mean airway pressure (MAP) (17.8 +/- 0.8 versus 15.6 +/- 0.5 cm H2O, p < 0.05) than CV. No significant difference in Pa(O2) and shunt fraction (QS/QT) was observed (83 +/- 7 mm Hg and 40 +/- 4% in CV versus 92 +/- 14 mm Hg and 35 +/- 3% in IRV, respectively). The Pa(CO2) was lower in IRV (48 +/- 3 versus 55 +/- 5 mm Hg, p < 0.05). Cardiac index (CI) and oxygen delivery (D(O2)) were lower in IRV (3.7 +/- 0.4 L/min/m2 and 500 +/- 61 ml/min/m2 versus 4.6 +/- 0.5 L/min/m2 and 617 +/- 80 ml/min/m2, respectively, p = 0.05 for both). Regardless of the considered parameter, no significant difference was observed between results after 1, 2, 4, and 6 h in each mode. We conclude that IRV at a ratio that results in a significant intrinsic PEEP does not improve Pa(O2), enhances CO2 elimination, decreases cardiac output (CO), and does not exert any time-dependent effect.


[Microcirculatory changes in infancy and old age].
An account of the microcirculatory unit according to Zweifach is followed by an explanation of the marked differences in capillary and tissue perfusion in children and aged. Prolonged hypertonia of the metarteriolar smooth muscle cells of the precapillary sphincters depresses capillary perfusion, even though the preferential circulation remains, resulting in marked changes in cellular metabolism in the tissues. If the capillary circulation is excluded for a sufficiently long period, less oxygen and substrates will be transported to the tissues concerned, intravasal red cell clumping will exacerbate hypoxia, and drainage of metabolic residues will be reduced. Microcirculatory vasoconstriction may be the result of heat, touch, noise, hormonal factors, medicaments, toxic, nervous, psychic factors, etc. Whit time, continuous damage to the microcirculation leads to involution of the microcirculation in favour of the preferential circulation, creation of an increasingly large number of arteriovenous short-circuits, and hyperstomia of arteriovenous anastomoses. Depressed capillary circulation leads to a reduced flow of liquids through the vessel walls owing to a diminished diffusion surface with consequent progressive lessening of tissue metabolic activity. Involution of the microcirculation is thus one of the main steps in the ageing process. A cell's metabolic pattern is dependent on the environment in which it lives, as well as its stock of enzymes and differentiation stage. This environment is greatly changed by alterations of the microcirculation. The gradual depression of the microcirculation during ageing, coupled with the consequent diminution of perfusion of tissues and interstitial liquids are thus of significance in ageing itself, in the slowing down of tissue change, and increased entropy to the energy levelling coincident with death.


Risk of pancreatic cancer in breast cancer families from the breast cancer family registry.
Increased risk of pancreatic cancer has been reported in breast cancer families carrying BRCA1and BRCA2 mutations; however, pancreatic cancer risk in mutation-negative (BRCAX) families has not been explored to date. The aim of this study was to estimate pancreatic cancer risk in high-risk breast cancer families according to the BRCA mutation status. A retrospective cohort analysis was applied to estimate standardized incidence ratios (SIR) for pancreatic cancer. A total of 5,799 families with ≥1 breast cancer case tested for mutations in BRCA1 and/or BRCA2 were eligible. Families were divided into four classes: BRCA1, BRCA2, BRCAX with ≥2 breast cancer diagnosed before age 50 (class 3), and the remaining BRCAX families (class 4). BRCA1 mutation carriers were at increased risk of pancreatic cancer [SIR = 4.11; 95% confidence interval (CI), 2.94-5.76] as were BRCA2 mutation carriers (SIR = 5.79; 95% CI, 4.28-7.84). BRCAX family members were also at increased pancreatic cancer risk, which did not appear to vary by number of members with early-onset breast cancer (SIR = 1.31; 95% CI, 1.06-1.63 for class 3 and SIR = 1.30; 95% CI, 1.13-1.49 for class 4). Germline mutations in BRCA1 and BRCA2 are associated with an increased risk of pancreatic cancer. Members of BRCAX families are also at increased risk of pancreatic cancer, pointing to the existence of other genetic factors that increase the risk of both pancreatic cancer and breast cancer. This study clarifies the relationship between familial breast cancer and pancreatic cancer. Given its high mortality, pancreatic cancer should be included in risk assessment in familial breast cancer counseling.


Prototyping context-aware nursing support mobile system.
A context aware nursing support system to push right information to the right person at the right moment is the key to increase clinical safety under a computerized hospital. We prototyped a system which obtains context from positions of nurses and list of expected clinical procedures. A WoZ test showed that the proposed approach has potential to decrease incidents caused by information delivery error.


Hierarchical Fullerene Assembly: Seeded Coprecipitation and Electric Field Directed Assembly.
Hierarchical C60 colloidal films are assembled from nanoscale to macroscale. Fullerene molecular crystals are grown via seeded cosolvent precipitation with mixed solvent [tetrahydronaphthalene (THN)/trimethylpyridine (TMP)] and antisolvent 2-propanol. The fullerene solutions are aged under illumination, which due to the presence of TMP reduces the free monomer concentration through fullerene aggregation into nanoparticles. The nanoparticles seed the growth of monodisperse fullerene colloids on injection into the antisolvent. Diverse colloidal morphologies are prepared as a function of injection volume and fullerene solution concentration. The high fullerene solubility of THN enables C60 colloids to be prepared in quantities sufficient for assembly (5 × 108 ). Electric fields are applied to colloidal C60 platelets confined to two dimensions. The particles assemble under dipolar forces, dielectrophoretic forces, and electrohydrodynamic flows. Frequency-dependent phase transitions occur at the critical Maxwell-Wagner crossover frequency, where the effective polarizability of the particles in the medium is substantially reduced. Structures form as a function of field strength, frequency, and confinement including hexagonal, oblique, string fluid, coexistent hexagonal-rhombic, and tetratic.


Leucine stimulates mammalian target of rapamycin signaling in C2C12 myoblasts in part through inhibition of adenosine monophosphate-activated protein kinase.
Mammalian target of rapamycin (mTOR) signaling is one of the main signaling pathways controlling protein synthesis. Leucine treatment upregulates mTOR signaling, which enhances protein synthesis; however, the mechanisms are not well understood. Herein, treatment of C2C12 myoblast cells with leucine enhanced the phosphorylation of mTOR and ribosomal protein S6 kinase. Leucine treatment also decreased the adenosine monophosphate/ATP ratio in myoblasts by 36.4 +/- 9.1% (P < 0.05) and reduced the phosphorylation of adenosine monophosphate-activated protein kinase (AMPK) alpha subunit at Thr172 (28.6 +/- 4.9% reduction, P < 0.05) and inhibited AMPK activity (43.6 +/- 3.5% reduction, P < 0.05). In addition, leucine increased the phosphorylation of mTOR at Ser2448 by 63.5 +/- 10.0% (P < 0.05) and protein synthesis by 30.6 +/- 6.1% (P < 0.05). Applying 5-aminoimidazole-4-carbox-amide 1-beta-d-ribonucleoside, an activator of AMPK, abolished the stimulation of mTOR signaling by leucine, showing that AMPK negatively controls mTOR signaling. To further show the role of AMPK in mTOR signaling, myoblasts expressing a dominant negative AMPKalpha subunit were employed. Negative myoblasts had very low AMPK activity. The activation of mTOR induced by leucine in these cells was abated, showing that AMPK contributed to mTOR activation. In conclusion, leucine stimulates mTOR signaling in part through AMPK inhibition. This study implicates AMPK as an important target for nutritional management to enhance mTOR signaling and protein synthesis in muscle cells, thereby increasing muscle growth.


[Suicidal behavior and the broken home. Critical evaluation of a "causal hypothesis"].
It has often been hypothesized that a broken home results in a "predisposition" towards suicidal behaviour. Previous findings concerning the incidence of broken homes among suicides have revealed rates that are considerably higher than might have been expected from the base rate of the incidence of broken homes among the population as a whole. However, when considering these results, it is necessary to take into account a number of general methodological problems as well as specific problems inherent in the study of the causes of suicidal behaviour. Therefore, due to methodological objections, it can be assumed that the covariation between the variable, broken home, and suicidal behaviour, as compared with other psychiatric disturbances, has been overestimated. It seems more likely that a broken home, in combination with the simultaneous and subsequent emergence of adverse circumstances, increases the probability of the appearance of deviant behaviour in general. Therefore, it would appear that the most useful approach for understanding suicidal behaviour is to analyse the learning history and background of patients who have exhibited such behaviour, while the variable, broken home, should be regarded as being only one of several factors conditioning the failure to develop or apply adequate coping strategies.


Subclassification of non-small cell lung carcinomas lacking morphologic differentiation on biopsy specimens: Utility of an immunohistochemical panel containing TTF-1, napsin A, p63, and CK5/6.
The availability of targeted therapies has created a need for precise subtyping of non-small cell lung carcinomas (NSCLCs). The aim of this study was to assess the utility of immunohistochemical markers in subtyping poorly differentiated NSCLC and to compare the results of immunohistochemical staining on biopsies with the corresponding resections. Thirty-nine cases of NSCLC that could not be further classified on biopsy and had subsequent resection specimens were identified. Classification of the tumor was based on the resection specimen using the World Health Organization criteria. All biopsies and resections were stained with CK7, TTF-1, napsin A (novel aspartic proteinase of the pepsin family), p63, CK5/6, and 34βE12. The specimens included 20 adenocarcinomas (ACs), 15 squamous cell carcinomas (SCCs), and 4 large-cell carcinomas (LCCs). TTF-1 was positive in biopsies from 16 of 20 ACs, 2 of 4 LCCs, and none of the SCCs. p63 was positive in all 15 SCCs, 2 of 20 ACs (both were also positive for TTF-1 and napsin A), and none of the LCCs. CK5/6 was positive in 11 of 15 SCCs (all p63 positive) but none of the ACs or LCCs. Napsin A stained 11 of 19 ACs (all TTF-1 positive) but none of the other tumors. Staining for CK7 was present in 19 of 19 ACs and 9 of 15 SCCs. 34βE12 stained both SCCs (15 of 15) and ACs (12 of 20). The combination of TTF-1, napsin A, p63, and CK5/6 allowed an accurate classification of 30 of39 (77%) cases. Of 232 pairs of slides (biopsy and resection) stained with immunohistochemical markers, 12 (5%) showed discrepancies in immunohistochemical staining between biopsies and their corresponding resections. Immunohistochemical staining using a combination of TTF-1, napsin A, p63, and CK5/6 allows subclassification of poorly differentiated NSCLCs on small lung biopsies in most cases. Discrepancies in immunohistochemical staining between biopsies and resections are uncommon.


Engineering complex phenotypes in industrial strains.
The global demand is rising for greener manufacturing processes that are cost-competitive and available in a timely manner. This has led to the development of a series of new tools and integrative platforms enabling rapid engineering of complex phenotypes in industrial microbes. By blending "old classical methods" of strain isolation with "newer approaches" of cell engineering, researchers are demonstrating the ability to stack multiple complex phenotypes in industrial hosts with some level of certainty. Newer tools for dissecting the genotype-phenotype correlation include association analysis (Precision Engineering), multiSCale Analysis of Library Enrichment (SCALE) in competition experiments, whole-genome transcriptional analysis, and proteomics and metabolomics technology. These newer and older tools of metabolic engineering and synthetic biology when combined with recent whole cell engineering approaches like whole genome shuffling, global transciptome machinery engineering, and directed evolutionary engineering, provide a powerful platform for engineering complex phenotypes in industrial strains. This review attempts to highlight and compare these newer tools and approaches with traditional strain isolation procedures as it applies to genome engineering with examples taken from literature.


Comparison of stainless steel stents coated with turbostratic carbon and uncoated stents for percutaneous coronary interventions.
Stent coating with turbostratic carbon was supposed to minimize the local inflammatory response after stent implantation and to thereby also reduce the rates of restenosis and clinical events. From October, 1999 to February, 2002, a total of 329 patients with symptomatic coronary artery disease (CAD) eligible for single-lesion PCI were randomized for implantation of either a CarboStent (C; n = 168) or a stainless-steel stent (S; n = 161). The stainless-steel stents were Tristar stents in 60.2%, Tetra stents in 17.4% and Penta stents in 22.4%. Both groups showed no differences in baseline clinical and angiographic characteristics. Angiographic follow-up (FU) was obtained after 6 months in 287 patients (87.2%), clinical FU in 295 patients (89.7%). With the exception of a smaller post-procedure minimal luminal diameter (MLD) in the C group (2.59 0.43 mm versus 2.72 0.46 mm in the S group; p = 0.01), there were no significant differences between the C and S groups in lesion length (10.28 4.45 mm versus 10.37 4.79 mm, respectively), reference diameter (2.92 0.59 mm versus 2.89 0.53 mm, respectively), pre-procedure MLD (0.77 0.36 mm versus 0.84 0.36 mm, respectively), MLD at FU (1.67 0.64 mm versus 1.68 0.57 mm, respectively), late loss (0.93 0.63 mm versus 1.05 0.59 mm, respectively), late loss index (0.51 0.32 versus 0.57 0.32, respectively) and restenosis rate (18.1% versus 20.6%, respectively). There were also no significant differences regarding major adverse cardiac events (MACE) between the C and S groups, i.e., occurrence of death (0% versus 0.7%, respectively), myocardial infarction (0% versus 0.7%, respectively), coronary artery bypass graft (0.7% versus 1.4%, respectively) and target lesion revascularization (16.4% versus 21.5%, respectively). Coronary stents coated with turbostratic carbon gave no clinically relevant reduction of in-stent restenosis and MACE rates when compared to uncoated stents.